Multiplex PCR-based synchronous and rapid method for detecting 13 pathogenic microorganisms in water

Abstract

The invention relates to a multiplex PCR-based synchronous and rapid method for detecting 13 pathogenic microorganisms in water, which comprises the steps of using multiplex PCR to simultaneously amplify gene-specific fragments of the 13 pathogenic microorganisms including escherichia coli, enterohaemorragic escherichia coli o157:h7, legionella pneumophila, salmonella enteritidis, shigella dysenteriae, staphyloccocus aureus, listeria monoeytogenes, helicobacter pylori, mycobacterium tuberculosis, klebsiella pneumonia, vibrio cholera, bacillus anthracis and yersinia pestis, and detecting PCR amplified products through agarose gel electrophoresis, thereby achieving synchronous and rapid detection for the 13 pathogenic microorganisms.

Description

technical field [0001] The invention belongs to the technical fields of environmental science and engineering and environmental protection. Background technique [0002] At present, the methods commonly used to detect and identify pathogenic microorganisms at home and abroad mainly include traditional biochemical identification methods, immunological detection techniques, conventional PCR and other simple molecular biology and immunological methods. [0003] The traditional biochemical identification method (such as isolation culture, biochemical identification, etc.) is the most widely used, and it is the national standard testing procedure such as "Food Sanitation Microbiological Inspection (GB / T 4789-2003)". This method can obtain the number and characteristics of bacteria in the sample, etc. Qualitative and quantitative results are widely used due to simple operation, economy and good accuracy, but most of them need to go through 4 steps of pre-enrichment, enrichment, se...

AI Technical Summary

Problems solved by technology

Although conventional PCR is a specific, sensitive, simple and rapid detection technology, it requires a long time for sample purification process, and once there is a very small amount of exogenous DNA contamination, false positive results may occur; multiple pairs of primers simultaneously Amplification, improper control of various experimental conditions can easily lead to amplification failure or non-specific products; improper primer design and target sequence selection may reduce its sensitivity and specificity; at the same time, generally only one pathogen can be detected at a time. Disease microorganism

Images